Isolation and formulations of nutrient-rich carotenoids

ABSTRACT

Pharmacologically and biologically active compositions containing carotenoids, in combination with micro and macro nutrients, a process for their preparation from carrots and their use in formulations for health care and nutrition applications. The process includes sequentially treating carrot juice with a carboxylic acid and a saccharide to obtain a carotenoid fraction rich in micro and macro nutrients in proportions compatible with those originally found in the natural state. A method of treating retenoid deficient states and immunomodulation is also disclosed using the composition.

This is a Division of Ser. No. 09/130,350 filed Aug. 4, 1998 now U.S.Pat. No. 6,056,962.

FIELD OF THE INVENTION

The invention relates to pharmacologically and biologically activecompositions containing carotenoids, micro and macro nutrients, aprocess for their preparation from carrots and their use in formulationsfor health care and nutrition applications.

BACKGROUND OF THE INVENTION

Carotenoids are a class of naturally-occuring yellow, orange or redtetraterpenoids, found in traces in plant tissue, algae, bacteria andfungi. In particular they are found in vegetable sources such ascarrots, spinach, tomatoes and fruits, such as, mango, peach, pumpkim,pappaya. The more commonly known carotenoids are, alpha-carotene,beta-carotene, lutein, zeaxanthin, lycopene and cryptoxanthin.

Carotenoids possess significant nutritional value, carotenes andcryptoxanthin being considered as a provitamin A precursor for theformation of retinal and Vitamin A in humans. Vitamin A, an essentialvitamin, for life is not synthesized in the animal cell.

Because carotenoids occur naturally in only trace amounts, thecarotenoids must be extracted in concentrated form in order to beuseful. Further carotenoids are sensitive to oxygen, air, heat andlight.

Nutritionists advocate the the daily use of carrots apart from otherfresh vegetables and fruits in diet. Carrots are grown seasonally andgood quality carrots are not available throughout the year at affordableprices. A little advertised fact about carrots is that only 20% of thetotal carotenes present are absorbed even when carrots are eaten in afinely grated form. The percentage of absorption from coarsely gratedraw or cooked carrots is still less being around 5%.

The low absorption is attributed to the poor permeability of thecellulosic cell wall to carotenes even after cooking with the resultthat the major part remains enclosed within the cells. Hence as a sourcefor deriving Vitamin A and use by themselves, carotenoid supplementsfree from cellulose are strongly recommended. Another advantage of usingprovitamin A carotenoids is that indiscriminate use of Vitamin A leadsto serious toxic effects (Hypervitaminosis) whereas even in large dosescarotenoids are harmless.

U.S. Pat. No. 2,567,362 describes fractional centrifugation process forthe separation of colloidal dispersoids of active plant pigment unitsfrom the generated vegetable hydrosol.

U.S. Pat. No. 2,739,145 describes the separation of coagulatedcarotenoid-protein particles following heating of a suspension of thevegetable fibre-separated particles in vegetable serum.

U.K. patent 776,405 describes a carotene-concentrate used as a foodstufffor animals using calcium hydroxide and subsequent pH adjustment withphosphoric acid and then with formic acid.

U.S. Pat. No. 2,848,508 describes a process for recovery of carotenefrom carrots and provides a saturated solution of carotene in naturalcarrot oil from carrots.

WTO 86/04059 describes the use of a pectolytic enzyme followed byultrafiltration for extracting and concentrating carotene.

U.S. Pat. No. 5,245,095 describes the use of calcium chloride, calciumhydroxide, calcium lactate or calcium gluconate to extract carotenoidsfrom natural sources.

None of the above or any known processes provide a compositioncontaining carotenoids in combination with micro and macro nutrients,the carotenoids being bound in moieties as in the natural source ofcarotenoids.

NATURE OF THE INVENTION

In recent times, carotenoids have been found in epidemiologicalobservations to display protective activity as physiologicalantioxidants, thus reducing the risks of development of several chronicdisorders such as heart diseases, cancer, cataract and other ailments.Holistic systems of medicine have, however, for centuries beenadvocating the synergistic value of dispensing not just pure but naturalproduct ingredients. Use of pharmacologically or biologically activeplant extracts is well-known.

Naturally occurring plant material contain a series of closely-relatedcompounds produced naturally via biological and biochemical reactions.The plant is capable of producing a wide range of analogues at least oneof which possesses the desired receptor compatibility. However, therelated compounds appear to exercise a synergistic effect on thepharmacological or biological activity of the compatible compound and atthe same time suppress toxic effects. Therefore the use of a compositeset of nutrients as they are present in the natural source, is rapidlygaining supporters in modern medicine.

However, a major drawback in using plant material in its crude form orto use the plant material in its natural state, is that the dosagesrequired of such material, to be therapeutically beneficial, are quitehigh. For example for receiving the therapeutic supply of carotenoidsabout one Kg of good quality carrots will have to be consumed every day.Such quantities cannot be conveniently converted into suitable dosageforms.

This invention, therefore, seeks to disclose a process for obtaining apharmacologically or biologically active plant extract substantially asit occurs in its natural state suitable for converting in a convenientadministrable dosage form. The process according to this invention,seeks to provide a concentrated plant extract, in which the plantextract comprises all pharmacologically or biologically active chemicalsin the proportions as they exist in the original natural state withoutthe use of organic solvents at any stage of its manufacture and avoidingthe use of oils, enzymes, mineral acids, alkalis, metal salts andtreatment by temperature in excess of 60 degrees celsius.

The advantage of naturally occurring series and analogues of compoundsis achieved without compromising the overall effects of these compounds.

There is no report to date of a product which is standardised in respectof both the carotenoids content as well as the micro and macro nutrientswith which the carotenoids are bound to and associated in its naturalstate occurring in vegetables, particularly carrots.

The present invention describes for the first time biologically orpharmacologically active material obtained from carrots, standardisedwith respect to its content of carotenoids, vitamins, proteins, lipids,carbohydrates, mineral and trace elements.

The present invention also describes a process to obtain the saidmaterial from carrots, which process comprises the addition of acarboxylic acid as herein defined to appropriately processed carrots,followed by the addition of a carbohydrate as herein defined, separationof a carotenoids-rich paste, and subsequent drying.

The present invention also describes the use of the biologically orpharmacologically active material in the preparation of formulations forhealth care and nutrition applications for use in as variety ofprophylactic and therapeutic conditions.

SUMMARY OF THE INVENTION

According to this invention there is provided a pharmacologically andbiologically active composition extracted from carrots, including0.25-5% mass as a percentage of total mass of extract of activecarotenoid fraction, absorbable by an animal or human body in aconvenient dosage form, in combination with micro and macro nutrientsaiding in the absorption, assimilation and supplementing the action ofthe carotenoid fraction.

Typically, the carotenoid fraction includes alpha-carotene,beta-carotene, lutein, zeaxanthin and lycopene.

Typically the micro nutrients are 0.01 to 1% vitamins, particularly theB complex vitamin, B1, B2, niacin and Vitamin C and 3 to 10% of mineralsand trace elements, as a percentage of total mass of extract.

Typically, the macro nutrients are 20-40% lipids, 10-50% proteins and 1to 25% carbohydrates as a percentage of total mass of extract.

The invention also provides a process for making a pharmacologically andbiologically active composition extracted from carrots comprising thesteps of

comminuting cleaned and washed carrots to obtain a homogeneouscomminution;

separating the juice from the comminution by filtration;

treating the juice with a carboxylic acid to adjust the pH of the juiceto between 3 and 6;

treating the pH adjusted juice with at least one saccharide;

centrifuging the saccharide containing juice to obtain the compositionin a paste form.

In accordance with another embodiment of this invention the processincludes a further step of drying the paste in vacuum and pulverizingthe solid material so formed to obtain the composition in particulateform.

Typically, the carboxylic acid is at least one acid selected from agroup consisting of mono carboxylic acid such as, ascorbic acid and/orsorbic acid, and/or a dicarboxylic acid such as adipic acid, malic acid,fumaric acid or tartaric acid or mixtures of them, and/or atricarboxylic acid such as citric acid, in solid form or as a saturatedaqueous solution in an amount of acid equivalent to 0.03-3.0% mass ofjuice.

Typically, the saccharide is at least one selected from a groupconsisting of monosaccharide such as fructose and/or dextrose, and/or adisaccharide such as sucrose, lactose, and/or hexitols such as mannitol,sorbitol, either in solid form or as a saturated aqueous solution, in anamount of saccharide ranging from 1-50% of the juice, preferably 20-30%of the mass of juice.

According to this invention there is further provided a method oftreating the human or animal body, therapeutically or prophylacticallyfor conditions arising from a retenoid deficiency state, oxidativestress, Wald's cycle aberration, pathological keratinzation,malignancies or for immunomodulation, by administering orally insuitable tablet, capsule or liquid dosage form or topically apharmaceutical or biological composition which contains, extracted fromcarrots, a 0.25 to 5% mass of active carotenoid fraction as a percentageof total mass of extract in combination with micro and macro nutrientsaiding in the absorption, assimilation and supplementing the action ofthe carotenoid fraction.

In accordance with one preferred embodiment of this invention the methodalso provides for including in the administered composition therapeuticamounts of at least one substance selected from a group containingspirulina, Vitamin E, Vitamin C, selenium compounds, zinccompounds,naturally occurring carotenoids such as those found in algae,fruits and vegetables.

In the sub therapeutic form the pharmacologically and biologicallyactive composition extracted from carrots can be used as a biocompatiblepigment.

DETAILED DESCRIPTION OF THE INVENTION

A principal object of the present invention is to provide apharmacologically and biologically active composition extracted fromcarrots (Daucus carota L), including 0.25-5% mass as a percentage oftotal mass of extract of active carotenoid fraction, absorbable by ananimal or human body in a convenient dosage form, in combination withmicro and macro nutrients aiding in the absorption, assimilation andsupplementing the action of the carotenoid fraction.

A feature of this invention is that the extracted composition iscompatible with the different natural constituents of the composition,namely the individual and total carotenoids, vitamins, proteins, lipids,carbohydrates, minerals and trace elements and other suchnaturally-occurring constituents as analyzed by methods known in theliterature. The amount of different naturally occurring constituents ofcarrots, is determined to lie within the ranges specified in thedescription below and in the accompanying examples. Generally forcarrots, the ranges per 100 grams of isolated powder is 250-5000 mgtotal carotenoids,is 10-1000 mg vitamins, 10-50 g proteins, 20-40 glipids, 1-25 g carbohydrates and 3-10 g minerals & trace elements.

A second object of the present invention is to describe a novel processfor the preparation of the composition from carrots. As one particularexample according to the invention, fresh, hard, good quality, orange orred colored carrots with a smooth surface are selected. Differentreddish varieties of carrots such as for example “Pusa Kesar”, “PusaMeghali”, “Desi Red” are available. Orange-coloured varieties such as“Bangalore local” and “Ooty Hybrid” are known to be rich inbeta-carotene, while the reddish-coloured varieties are more rich inlycopene. The carrots are thus selected for process depending on thenature of the composition to be prepared. Defective carrots areeliminated or excluded in the sorting out process. The selected carrotsare washed thoroughly with water and comminuted in an appropriate mill,typically a fruit mill which consists of a rotating stainless steelblades whose speed of rotation is adjustable to between 100 to 1000 rpmand which is fitted with a sieve with apertures variable from 1 to 10 mmto control the particle size of the homogeneous comminution. Thecomminution is treated through a filter press or a coarse filter (50-150microns) for the purpose of separating the pulp from the juice. To thejuice is added with stirring a mono-carboxylic acid such as ascorbicacid, and/or sorbic acid, and/or a dicarboxylic acid such as adipicacid, malic acid, fumaric acid or tartaric acid or mixtures of them,and/or a tricarboxylic acid such as citric acid, in solid form or as asaturated aqueous solution in an amount of acid equivalent to 0.03-3.0%of the liquid, such that a pH value of the resulting mixture of about3.0-6.0, preferably 5.0 is reached. This helps to build up the particlesize of colloidal carotenoid complexes enabling the further processingof the juice by filtration or centrifuging. The carboxylic acids alsostabilise the juice during processing. To the resulting solution orsuspension is added one or more components of a monosaccharide such asfructose and/or dextrose, and/or a disaccharide such as sucrose,lactose, and/or hexitols such as mannitol, sorbitol, either in solidform or as a saturated aqueous solution, in an amount of saccharideranging from 1-50% of the juice, in particular 20-30% of the carrotjuice, which mixture is subjected to centrifugation to providecarotenoids-rich carrot paste containing 0.1-1.0% carotenoids. It hasbeen observed that the use of saccharides not only increases thestability of the carotenoid fraction but also is instrumental inextracting the carotenoid fraction bound to lipoproteins in combinationwith micronutrients such as the B-complex vitamins and minerals. In theabsence of the saccharides step, the concentration of micro nutrients inthe final composition is less than 50 per cent. Moreover, in the absenceof the above, the carotenoids degrade within a few months storage. Thecarotenoids-rich paste can be used as such or after drying under highvacuum, pulverizing and sieving through appropriately sized sieves bywhich the pharmacologically and biologically active composition of theinvention is obtained in powder form.

The powder is analyzed according to known procedures to provide theprecise composition of the carotenoids, vitamins, minerals & traceelements, proteins, carbohydrates and lipids in the powder.

The present invention also describes the use of the paste or powder ofthe invention in health care and nutrition applications and as acolouring agent. The paste can be formulated as an emulsion orsuspension for oral or topical use or as a colouring matter usingappropriate excipients and adjuvants known to those skilled in theformulations art. The powder can be formulated as solid dosage forms fororal use as powder/granules or as capsules/tablets. These can also becombined with other antioxidants, minerals, vitamins and othermicronutrients. Carotenoids act as lipid phase antioxidants. Howeverbeta-carotene supplementation alone does not appear to reduce thesusceptibility of LDL to oxidation. The present invention also disclosesa method of treating the human or animal body, therapeutically orprophylactically for conditions arising from a retenoid deficiencystate, oxidative stress, Wald's cycle aberration, pathologicalkeratinzation, malignancies or for immunomodulation, LDL cholesterolreduction, cancer adjuvant therapy and reducing risk of cardiovasculardisease by administering orally in suitable tablet, capsule or liquiddosage form or topically a pharmaceutical or biological compositionwhich contain extracted form carrots, a 0.25 to 5% mass of activecarotenoid fraction as a percentage of total mass of extract incombination with micro and macro nutrients aiding in the absorption andassimilation and supplementing the action of the carotenoid fraction.Inflammatory and allergic manifestations in the living cell are thoughtto be the direct cause of hyperactivity of immune function entities innon-specific immunity, whereas the suppression or deficiency of immunefunctions are the result of hypoactivity. The functioning and efficiencyof non-specific immunity may be influenced by many exogenous andendogeneous factors like physical and psychological, oxidant orhyperoxidative stress, hormonal imbalance, pharmaceuticals and the like.A recent trend in medicine is to consider all disease from the molecularperspective, in which derangements in the structures or conformations ofvital biomolecules in diseased states are intricately implicated in theaetiopathogenesis of those diseased conditions.

‘Immunomodulation’ is any procedure which can alter non specificimmunity by interfering with its functioning. If it results inenhancement of immune reactions, it results in immunostimulation andprimarily implies stimulation of the non specific immunity, that isstimulation of the function and efficiency of granulocytes, macrophages,natural killer cells, complement and properdin, and the various effectorsubstances including interleukins, tumour necrosis factor, interferons,lysozymes, prostaglandins, oxygen radicals and other mediators.Immunosuppression mainly implies reduced resistance against infectionsand stress and may be due to environmental or chemotherapeutic factors.

Immunostimulation and immunosuppression both need to be addressed inorder to regulate normal immunological functioning. Hence,immunostimulating and immunosuppressing agents both have their ownstanding. There are a variety of known immunosuppressing agents, forinstance cyclosporin, however few immunostimulating agents areavailable. Apart from specific stimulative or suppressive activity, itis believed that certain agents of plant origin such as the carotenoidshave the activity to normalize or modulate pathophysiological processesin the underlying immune response and hence the term immunomodulation orimmunomodulatory agents or adaptogenic agents are used for these agents.This activity is believed to be dose dependant as can be seen from theimmunostimulation at low dilutions of many immunosuppressants. Thus in abiological system, active material will act as an immunostimulant in lowdoses but as an immunosuppresant in high doses. Such a biologicallyactive material can be called as an ‘immunomodulator’.

Biological membranes contain phospholipids (PLs) and degradation ofthese by oxygen can cause loss of cell integrity. PLs contain highquantities of polyunsaturated fatty acids (PUFAs) and the double bondsof these fatty acids are easily attacked by oxygen to produce toxicfatty peroxides. The extent of formation of peroxides (peroxidation)increases with the number of double bonds in the fatty acids of the PLs,so membranes with a high PUFA content are specially likely to be moreoxidized. A common example of peroxidation is rancidity of butter andvegetable oils. Another agent produced within and without the cell iscalled a free radical. These substances are highly reactive because theyare chemically incomplete and hence unstable, so they can latch on toother substances very readily. They are also known as super oxideradicals (singlet oxygen) and they are produced within the cells both byby self-oxidation (as in peroxides) and by enzymatic processes. Theirhigh intrinsic reactivity and their ability to generate even more potentoxidizing agents when combined with peroxides constitutes a constantthreat to cellular integrity. It must be admitted that free radicalperform some useful functions: the bactericidal action of leucocytes andin mediating inflammatory responses are notable examples, but it is whenthey are produced in large quantities and their metabolic products areallowed to go unchecked that they can seriously damage membranes andeven denature DNA. Increased generation of free radicals in thebiological systems over and above the potential of antioxidantmechanisms to curb them produces a state what is commonly known asoxidative (oxidant) stress in the body. Since free radicals arecontinuously being generated, the human body has developed a number ofmechanisms to deal with their potentially damaging effects and those oftheir metabolites. The susceptibility of any tissue to an oxidativestress induced by free radicals or peroxide relates to the balancebetween the extent of that stress and the antioxidant ability of theprotective agents present. ‘Health’ can be defined as an equilibriumstate between the generation and scavenging of free radicals andperoxides. The cellular defense mechanisms and scavenging agents involvevarious enzymes such as superoxide dismutase, glutathione synthetase,glutathione peroxida6e, glutathione reductase, glucose-6-phosphate,dehydrogenase and catalase. Plasma proteins with antioxidant potentialinclude the copper containing transferrin, ceruloplasmin and the ironcontaining transferrin. Foods constituents that also contribute toprotection include the sulphur containing amino-acids and the mineralsselenium, zinc and copper. In most pathological conditions theprotective antioxidant mechanisms are overwhelmed leading to an elevatedor rising state of free radicals and peroxides. Both Vitamin E andcarotenoids can successfully deactivate super oxides (singlet oxygen)but when Vitamin E attacks the oxide it is destroyed. On the other handbeta-carotene can quench the oxide without damage to itself and thus canbe used again and again and can convert singlet oxygen back to normaloxygen before it can cause damage leading to skin or lung cancer.Beta-carotene is also very efficient at trapping the free radicals.

Hence the present invention also encompasses that in the process of thepreparation of formulations, there may be added adjuvants to thecomposition of the invention, such adjuvants being Vitamin C, Vitamin E,Compounds of selenium such as Selenium Dioxide and compounds of zincsuch as Zinc Sulphate and additional amounts of commercially availablenatural-sourced carotenoids such as from fruits, vegetables and algae.The composition of typical preparations are described under respectiveexamples.

The formulations of the invention are useful for the prophylactic ortherapeutic treatment of subjects diagnosed to be carotenoid-deficientor Vitamin A-deficient or in need of protective efficacy of antioxidantsknown to be implicated in chronic disorders such as heart diseases,cancer, cataract and other chronic ailments. Both human and veterinaryuse is envisaged. The dosage per day is variable dependent on the ageand weight of the subject to be treated and the severity of thecondition as assessed by practicing medical physicians.

The following examples illustrate but do not limit the scope of theinvention.

EXAMPLE 1

Fresh, hard, good quality reddish colored “Desi Red” carrots with asmooth surface, excluding those that were found defective, were selectedand washed thoroughly with water. The sorted and washed carrots (1.0 kg)were subjected to crushing in a fruit mill to provide a comminutionwhich was subjected to pressing through a filter press for the purposeof separating the pulp from the juice to provide a juice (ca. 600 ml).To the juice, 3 g of adipic acid was added with stirring. To theresulting mixture was added 60 g of sorbitol and the mixture wassubjected to centrifuging to provide paste (ca. 17.2 g). The paste wasdried under high vacuum. Pulverizing of the solid material and sievinggave the carotenoid powder of the invention (3.8 g). The composition ofthe product is given below.

Composition per 100 g Product from “Desi Red” Carrots (Example 1)

beta-Carotene 530 mg alpha-Carotene 27 mg Lycopene 700 mgLutein/Xeaxanthin 15 mg Total Carotenoids 3750 mg Proteins 32.8 gCarbohydrates 4 g Phosphorus 647 mg Lipids 15.3 g Vitamin C 22 mgVitamin B1 5 mg Vitamin B2 1 mg Iron 95 mg Zinc 1 mg Manganese 1 mgMagnesium 162 mg Calcium 1.381 g Potassium 1.99 g Sodium 1.99 g TotalMinerals (Ash value) 6.87 g

EXAMPLE 2

The “Bangalore local” carrots were processed according to the proceduredescribed in Example 1. The composition of the product is given below.

Composition per 100 g Product from “Bangalore local” Carrots

beta-Carotene 2095 mg alpha-Carotene 31 mg Lycopene 70 mgLutein/Xeaxanthin 13 mg Total Carotenoids 2948 mg Phosphorus 680 mgProteins 31.5 g Carbohydrates 4.5 g Lipids 30.3 g Vitamin C 24 mgVitamin B1 6 mg Vitamin B2 2 mg Iron 12 mg Zinc 1.4 mg Manganese 1 mgMagnesium 602 mg Calcium 894 mg Potassium 2 g Sodium 1 g Total Minerals(Ash value) 7.1 g

EXAMPLE 3

The different supplies carrots were processed according to theprocedures described in Examples 1 to 13. The composition of the powderwas analyzed and shown to have values in the range as described below.

Range of composition per 100 g products from carrots (Daucus carota L)

beta-Carotene  100-4000 mg alpha-Carotene  10-300 mg Lycopene  10-2000mg Lutein/Xeaxanthin  5-50 mg Total Carotenoids  250-5000 mg Proteins10-50 g Carbohydrates  1-25 Phosphorus 0.1-1   g Lipids 20-40 g VitaminC  10-500 mg Vitamin B1 1-6 mg Vitamin B2 0.5-4   mg Iron  5-100 mg Zinc1-5 mg Manganese 0.1-1   mg Magnesium  50-900 mg Calcium 0.5-3   gPotassium 1-4 g Sodium 1-3 g Total Minerals (Ash value)  3-10 g

EXAMPLE 4

The procedure described in Example 1 was followed using citric acid inplace of adipic acid.

EXAMPLE 5

The procedure described in Example 1 was followed using fumaric acid inplace of adipic acid.

EXAMPLE 6

The procedure described in Example 1 was followed using malic acid inplace of adipic acid.

EXAMPLE 7

The procedure described in Example 1 was followed using tartaric acid inplace of adipic acid.

EXAMPLE 8

The procedure described in Example 1 was followed using ascorbic acid inplace of adipic acid.

EXAMPLE 9

The procedure described in Example 1 was followed using sorbic acid inplace of adipic acid.

EXAMPLE 10

The procedure described in Example 1 was followed using mannitol inplace of sorbitol.

EXAMPLE 11

The procedure described in Example 1 was followed using sucrose in placeof sorbitol.

EXAMPLE 12

The procedure described in Example 1 was followed using lactose in placeof sorbitol.

EXAMPLE 13

The procedure described in Example 1 was followed using dextrose inplace of sorbitol.

EXAMPLE 14

Preparation of nutrient-rich carotenoids tablets.

Typical composition of Ingredients:

Nutrient-rich carotenoid powder (cf Example 1) 400 mg Gelatin 12 mgSucrose 25 mg Microcrystalline Cellulose 25 mg Starch 60 mg Talc 5 mgMagnesium stearate 3 mg Colloidal Silicon dioxide 10 mg Hydroxy ProphylMethyl Cellulose 15 mg Titanium Dioxide 0.5 mg Sunset Yellow FCF 4 mgPropylene Glycol 1 mg

The tablets were prepared by blending nutrient-rich carotenoid powderwith sucrose and Microcrystalline cellulose, granulating with StarchGelatin paste, drying, lubricating with Talc, Magnesium stearate andColloidal silicon dioxide followed by compression into tablets. Forcoating, Hydroxypropyl Methyl Cellulose, Propylene glycol and a blend ofTitanium dioxide/Sunset Yellow FCF were used.

EXAMPLE 15

Preparation of nutrient-rich carotenoid in combination with otherantioxidants in tablet form.

Typical composition of ingredients:

Nutrient-rich carotenoid powder (cf Example 1) 100 mg Naturalbeta-carotene and carotenoids 20% 15 mg Vitamin E Acetate 25 mg VitaminC 150 mg Selenium dioxide 75 mcg Zinc, sulphate 70 mg MicrocrystallineCellulose 25 mg Starch 25 mg Gelatine 5 mg Talc 6 mg Magnesium Stearate4 mg Colloidal Silicon Dioxide 6 mg Hydroxypropyl Methyl Cellulose 15 mgTitanium Dioxide and Sunset Yellow FCF 0.5 mg Propylene Glycol 1 mg

Tablets were prepared by blending nutrient-rich carotenoid powder withVitamin E acetate, Vitamin C, Selenium Dioxide and Zinc Sulphate. Theblend was mixed with Microcrystalline Cellulose, granulated with Starchpaste, dried, lubricated with Talc, Magnesium stearate and ColloidalSilicon Dioxide followed by compression into tablets. For coating,Hydropropyl Methyl Cellulose, Propylene glycol Titanium Dioxide andSunset yellow FCF were used.

EXAMPLE 16

Preparation of tablets of nutrient-rich carotenoid with spirulina:typical composition of ingredients

Nutrient-rich carotenoid Powder (ef example 1) 250 mg Spirulina 250 mgMicrocrystalline Cellulose 50 mg Starch 15 mg Talc 6 mg MagnesiumStearate 4 mg Colloidal Silicon Dioxide 6 mg

The tablets were prepared by blending nutrient-rich carotenoid powder,Spirulina and Microcrystalline Cellulose, granulated with starch paste,dried, lubricated with Talc, Magnesium Stearate and Colloidal SiliconDioxide and compressed into tablets.

EXAMPLE 17

Preparation of nutrient-rich carotenoid capsules

Typical composition of ingredients

Nutrient-rich carotenoid powder (cf example 1) 250 mg MicrocrystallineCellulose 100 mg Talc 7 mg Magnesium Stearate 2 mg Colloidal SiliconDioxide 4 mg

Nutrient-rich carotenoid powder was blended with MicrocrystallineCellulose, lubricated with Talc, Magnesium Stearate, Colloidal silicondioxide and filled in the capsules.

EXAMPLE 18

Preparation of nutrient-rich carotenoid Soft Gelatin Capsules

Typical composition of ingredients

Nutrient-rich carotenoid powder (cf example 1) 250 mg Vegetable oil 250mg Soft gelatin capsule shell one

EXAMPLE 19

Preparation of nutrient-rich carotenoid powder mix

Typical composition of ingredients

Nutrient-rich carotenoid powder (cf example 1) 500 mg Sucrose 500 mg

Nutrient-rich carotenoid powder was blended with sucrose and filled in40 micron Aluminium-poly sachets.

EXAMPLE 20

Preparation of suspension of micronutrient-rich carotenoid paste

Typical composition of ingredients

Nutrient-rich carotenoid paste (cf Example 1) 20 g Sucrose 60 g Purifiedwater to make 100 ml

Nutrient-rich carotenoid paste was suspended in syrup prepared fromsucrose and water.

EXAMPLE 21

Preparation of cream containing nutrient-rich carotenoid paste

Typical composition of ingredients

Nutrient-rich carotenoid paste (cf Example 1) 1 g Vitamin E Acetate 1 gVitamin C 1 g Light Liquid Paraffin 10 g Propylene Glycol 8 gCetostearyl alcohol 7 g Cetomacrogol - 1000 3.5 g White Bees Wax 2.5 gPurified Water 66 g

A blend of nutrient-rich carotenoid paste, Vitamin E acetate and VitaminC was incorporated in cream prepared from solution of Cetostearylalcohol, Cetomacrogol 1000 and White beeswax in light liquid paraffinand water-propylene glycol mixture.

Tablets(Example 14) containing 400 mg of the composition equivalent to1.2 mg of carotenoid were used for trials:

GROUP I

5 participants complaining of watering of eyes, redness and inability toopen the eyes completely in sunlight were administered the tablets overa period of two weeks. Significant reduction of the symptoms wereobserved.

GROUP II

5 participants sensitive to continuous exposure to sunlight andsuffering from skin erythema on longer exposure were administered thetablets over a period of four weeks. Significant reduction of thesymptoms induced by long exposure were observed.

We claim:
 1. A method of inducing antioxidant activity in warm-bloodedanimals comprising administering to warm-blooded animals anantioxidatingly effective amount of a biologically active compositioncomprising 0.25 to 5% by weight of an active carotenoid fractionextracted from carrots with a carboxylic acid at a pH of 3 to 6 followedby treatment with at least one saccharide based on the total weight ofthe composition and micro and macro nutrients sufficient to aid in theabsorption acid assimilation of the carotenoid fraction and tosupplement the action thereof.
 2. The method of claim 1 wherein thecomposition also contains therapeutic amounts of at least one member ofthe group consisting of spirulina, Vitamin E, selenium compounds, zinccompounds, Vitamin C and carotenoids extracted from other naturalsources.
 3. The method of claim 1 wherein the macro nutrients include 20to 40% by weight of the composition of lipids.
 4. The method of claim 1wherein the macro nutrients include 10 to 50% by weight of the totalcomposition of proteins.
 5. The method of claim 1 wherein the macronutrients include 1 to 25% by weight of the total composition ofcarbohydrates.
 6. The method of claim 1 wherein the micro nutrientsinclude 0.01 to 1% by weight of the total composition of at least onevitamin selected from the group consisting of Vitamins B₁ and B₁, Niacinand Vitamin C.
 7. The method of claim 1 wherein the micro nutrientsinclude 3 to 10% by weight of the total composition of materials andtrace elements.
 8. The method of claim 1 wherein the carotenoid fractionincludes α-carotene, β-carotene, Lutein, zeaxanthin and lycopene.